Role of Efflux Pump Gene adeIJK to Multidrug Resistance in Acinetobacter baumannii Clinical Isolates

Ji Ae Choi; Choon-Mee Kim; Sook-Jin Jang; Seong-Sik Cho; Chul Ho Jang; Young-Jin Ko; Seongho Kang; Geon Park

doi:10.5145/ACM.2020.23.1.45

Journal List > Ann Clin Microbiol > v.23(1) > 1144295

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Choi, Kim, Jang, Cho, Jang, Ko, Kang, and Park: Role of Efflux Pump Gene adeIJK to Multidrug Resistance in Acinetobacter baumannii Clinical Isolates

Original Article

Ann Clin Microbiol 2020;23(1):45-54.

Published online: 12 March 2020

DOI: https://doi.org/10.5145/ACM.2020.23.1.45

Role of Efflux Pump Gene adeIJK to Multidrug Resistance in Acinetobacter baumannii Clinical Isolates

Ji Ae Choi¹, Choon-Mee Kim², Sook-Jin Jang³, Seong-Sik Cho⁴, Chul Ho Jang⁵, Young-Jin Ko³, Seongho Kang³, Geon Park³

¹Division of Antimicrobial Resistance, Center for Infectious Diseases Research, Korea National Institute of Health, KCDC, Cheongju

²Premedical Science, College of Medicine, Gwangju

³Department of Laboratory Medicine, College of Medicine, Gwangju

⁴Department of Laboratory Medicine, Chosun University Hospital, Gwangju

⁵Department of Otolaryngology, Chonnam National University Medical School, Gwangju, Republic of Korea

Corresponding author Sook-Jin Jang E-mail: sjbjang@chosun.ac.kr Tel: +82-62-220-3272 Fax: +82-62-232-2063

Received 20 September 2019 Revised 18 November 2019 Accepted 1 December 2019

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/4.0) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

Abstract

Background

The emergence of multidrug-resistant Acinetobacter baumannii as a nosocomial pathogen is one of the major public health problems. The aim of this study was to evaluate the role of an efflux pump gene adeJ for the multidrug resistance ofA. baumannii clinical isolates.

Methods

Two groups (MDRAB and SAB) of A. baumannii clinical isolates were studied. The SAB group consisted of strains that did not meet the criteria of MDRAB and were susceptible to more categories of antibiotics than MDRAB. Antimicrobial susceptibility results obtained by VITEKⅡ system were used in data analysis and bacterial group allocation. We performed real-time reverse transcription PCR to determine relative expression ofadeJ. We compared relative expression of adeJ in comparison groups by considering two viewpoints: i) MDRAB and SAB groups and ii) susceptible and non-susceptible groups for each antibiotic used in this study.

Results

The mean value of relative expression of adeJ of MDRAB and SAB groups was 1.4 and 0.92, respectively, and showed significant difference (P=0.002). The mean values of relative expression of adeJ of susceptible and non-susceptible groups to the antibiotics cefepime, ceftazidime, ciprofloxacin, imipenem, meropenem, tigecycline, piperacillin/ tazobactam, ticarcillin/clavulanic acid, trimethoprim/sulfamethoxazole, piperacillin, and gentamicin showed statistically significant differences.

Conclusion

The overexpression ofadeIJK might contribute to the multidrug resistance inA. baumannii clinical isolates. Further, the overexpression ofadeIJK might be one of the factors contributing to the resistance to numerous antibiotics.

Keywords: Acinetobacter baumannii, adeJ gene, Efflux, Multidrug resistance

References

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Fig. 1.

Relative expression of the adeJ efflux pump gene determined by real-time reverse transcriptase PCR in A. baumannii type strain (ATCC19606), multidrug resistant A. baumannii AYE strain, 70 strains of multidrug resistant A. baumannii (MDRAB) and 30 strains of non-MDRAB clinical isolates. The normalized expression of adeJ was calibrated with the expression of A. baumannii ATCC 19606. The error bars represent the standard error of the mean. *P < 0.05.

Fig. 2.

Relationship between adeJ pump gene expression and resistance to each antibiotics. Relative expression of adeJ efflux pump gene in non-susceptible (NS) group was higher than that of susceptible (S) group to each antibiotics in general. The statistically significant difference was found between S and NS groups to cefepime, cefotaxime, ceftazidime, ciprofloxacin, gentamicin, imipenem, meropenem, piperacillin, piperacillin/tazobactam, ticarcillin/clavulanic acid, tigecycline, and trimethoprim/sulfamethoxazole, respectively. *P < 0.05, **P < 0.001. The error bars represent the standard error of the mean.

Table 1.

Oligonucleotide primers used in this study

PCR	Gene	Primer	Sequences (5' → 3')	AT	References
Conventional PCR	gyrB Multiplex 1	Sp4F	CAC GCC GTA AGA GTG CAT TA	60°C	(11)
		Sp4R	AAC GGA GCT TGT CAG GGT TA
		Sp2F	GTT CCT GAT CCG AAA TTC TCG
	bla _OXA51-like	Oxa51-like F	TAA TGC TTT GAT CGG CCT TG	55°C	(12)
		Oxa51-like R	TGG ATT GCA CTT CAT CTT GG
Real-time RT-PCR	rpoB	rpoB F	CTC ACT ATG GTC GTG TTT GTC	57°C	this study
		rpoB R	CCA AGA AAC CGA AGT CAT TCG
	adeJ	adeJ F	CAA GTT ATT GCA TTC TAT TCA CCA G	57°C	this study
		adeJ R	GAC CTG TAC CTC ACC AAC AC

Abbreviation: AT, annealing temperature.

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